华南农业大学学报2017，Vol.38Issue(1)：15-22,8.DOI:10.7671/j.issn.1001-411X.2017.01.004
玉米淀粉合成酶SSⅡa启动子的克隆及功能分析
Cloning and function analysis of starch synthase SSⅡa promoter in maize
摘要
Abstract
Objective] To clone maize ( Zea mays) starch synthase SSⅡa promoter, analyze its function, and provide a basis for its future research and application. [Method] The SSⅡa 5′flanking sequence was found on Maize GDB by BLASTing the maize genome sequence published on NCBI, and the SSⅡa promoter was cloned from maize B73 using PCR. We analyzed the cis elements of the promoter using PlantCare. Fragments of 1 407, 867, 633, 483, and 365 bp were cloned with specific primers, and were inserted into the plant expression vector pCAMBIA3301, respectively. Five plant expression vectors with different 5′deletions of the SSⅡa promoter were constructed and named P1 , P2 , P3 , P4 and P5 . The transgenic Arabidopsis thaliana plants were obtained through Agrobacterium-mediated transformation.[Result] A DNA fragment of 2 526 bp was obtained by PCR amplification with maize B73 genome DNA as template and SSⅡaF/SSⅡaR as specific primers. The positive A. thaliana plants, which were screened by herbicide, had gus gene by PCR detection. The histochemical analysis of GUS showed that the expression vectors of five promoters were blue in leaves and pods at maturity. The quantitative analysis of gus gene showed that among five transgenic A. thaliana at maturity, the expression level of P1 in leaves was the highest and the others were basically the same, and the expression levels of P1 and P2 in seeds were similar, both being higher than those of P3, P4 and P5. [Conclusion] The maize SSⅡa promoter has been successfully cloned. The five constructed expression vectors with different 5′deletions of the SSⅡa promoter all have activities in transgenic A. thaliana, and the promoters with the length of 1 407 bp (P1) and 867 bp (P2) have endosperm specificity.关键词
玉米/淀粉合成酶/SSⅡa启动子/克隆/载体构建/功能分析Key words
maize/starch synthesis enzyme/SSⅡa promoter/cloning/vector construction/function analysis分类
农业科技引用本文复制引用
陈展宇,王阔,王晓梅,刘相国,崔喜艳..玉米淀粉合成酶SSⅡa启动子的克隆及功能分析[J].华南农业大学学报,2017,38(1):15-22,8.基金项目
国家自然科学基金(31200611) （31200611）
吉林省应用基础研究重点实验室开放课题(20130102066JC) （20130102066JC）
转基因生物新品种培育重大专项(2014ZX0801004B) （2014ZX0801004B）
